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To assess sarcoma margins with more accuracy and aid surgical planning, we constructed three-dimensional (3D) digital models with computed tomography(CT) and magnetic resonance imaging (MRI) image fusion data and validated the preciseness of the models by comparing them with 3D models constructed with CT only data.We retrospectively reviewed a consecutive set of patients treated in our center who were preoperatively evaluated with the fusion image model. Models based on fusion images or CT-only data were constructed. Volumes of both tumors were calculated and the tumors were overlapped to see the location of differences between the two models.A consecutive 12 cases (4 male vs. 8 female) were included in this study. Most of the tumors were located in the pelvic bone or spine. The volume of the two tumor models was different and the differences were mainly in the peripheral region of the tumor.CT and MRI fusion image 3D models are more accurate than models with CT-only data and can be very helpful in preoperative planning of sarcoma patients.

Gelatin methacrylate (GelMA) hydrogels are a promising material for use in a variety of tissue engineering applications. Herein, we focused on identifying the optimal irradiation time necessary to photopolymerize GelMA hydrogels with visible blue light in a manner that did not adversely impact the biophysical properties of these cell-containing gels. We assessed the toxic effects of different irradiation times (3, 5, 10, 20 and 40 seconds) on BMMSCs encapsulated in a GelMA hydrogel using lithium phenyl-2,4,6 trimethylbenzoylphosphinate (LAP) as a photoinitiator. Both CCK-8 assays and Live-Dead staining were used to measure BMMSCs viability. We observed increasing compression strength as a function of increased irradiation time, although this corresponded to a reduction in swelling ratio and pore sizes. We ultimately found that when using LAP as a photoinitiator, the optimal irradiation time was 5–10 seconds, which was suitable for bone tissue engineering application. Ultimately we determined that a 5 second irradiation time was optimal for studies of encapsulated stem cells.

Summary Aberrant RAS signaling activation is common in cancers with even few Ras mutations, indicating alternative dysregulation other than genetic mutations. We identified a Ras GTPase-activating gene RASA5/SYNGAP1, at the common 6p21.3 deletion, methylated/downregulated in multiple carcinomas and different from other RASA family members (RASA1–RASA4), indicating its special functions in tumorigenesis. RASA5 mutations are rare, unlike other RASA members, whereas its promoter CpG methylation is frequent in multiple cancer cell lines and primary carcinomas and associated with patient’s poor survival. RASA5 expression inhibited tumor cell migration/invasion and growth in mouse model, functioning as a tumor suppressor. RASA5 suppressed RAS signaling, depending on its Ras GTPase-activating protein catalytic activity, which could be counteracted by oncogenic HRas Q61L mutant. RASA5 knockdown enhanced Ras signaling to promote tumor cell growth. RASA5 also inhibited epithelial–mesenchymal transition (EMT) through regulating actin reorganization. Thus, epigenetic inactivation of RASA5 contributing to hyperactive RAS signaling is involved in Ras-driven human oncogenesis., Graphical Abstract, Highlights • RASA5 is normally widely expressed but epigenetically silenced in multiple cancers • Epigenetic disruption of RASA5 is associated with tumor progression in patients • RASA5 suppresses RAS signaling, depending on its RasGAP catalytic activity • RASA5 functions as a tumor suppressor through inhibiting EMT and metastasis, Biological Sciences; Molecular Biology; Cell Biology; Cancer

Purpose: Taurine has long been thought to be involved in retinal protection from retinal degenerative diseases, but the underlying molecular mechanisms remain unclear. Retinal neurodegeneration is an early event in the pathogenesis of diabetic retinopathy (DR) that precedes and participates in the microcirculatory abnormalities that occur in DR. Our objective was to investigate the role and mechanisms of taurine in early diabetic retinas.Methods: Eight-week-old STZ-induced diabetic rats and control animals were randomly assigned to receive taurine or vehicle by intraperitoneal injection or by intragastric administration. The retinal function and retinal cell counts were evaluated using an electroretinography (ERG) and immunofluorescence microscopy. Plasma amino acids were measured by ion-exchange chromatography (IEC). The expression levels of retinal taurine transporter (Tau-T), mitochondria-dependent apoptosis-associated genes and reactive gliosis markers were studied by western blotting and immunofluorescence. Pre- and post-synaptic markers (PSD95 and mGluR6) in outer plexiform layer (OPL), and the bipolar cell marker protein kinase C alpha (PKCα) were localized by immunofluorescence. Levels of PSD95 and mGluR6 were determined by quantitative western blot.Results: Taurine significantly prevented the reduction of photopic b-wave amplitude and retinal cone cells and ganglion cells loss and maintained the Bcl-2/Bax ratio balance in diabetic rats. Taurine also prevented the upregulation of glial fibrillary acidic protein (GFAP) and reduced retinal reactive gliosis. Taurine reduced plasma glutamate and tyrosine levels, which were elevated in diabetic rats. Moreover, mGluR6 levels reduction detected by western blot and immunofluorescence in diabetic retinas was inhibited and the displacement of mGluR6 in OPL into the inner nuclear layer (INL) detected by immunofluorescence was reduced by Taurine treatment.Conclusion: Taurine may protect retinal cells from diabetic attacks by activating Tau-T, reducing retinal reactive gliosis, improving retinal synaptic connections and decreasing retinal cell apoptosis. Thus, taurine treatment may be a novel approach for early DR.

This paper presents a geomagnetic field positioning model based on Bayes classifier. First, use the geomagnetic field sensor collecting geomagnetic field data, and establish a fingerprint database according to their statistical characteristics. Then, in the process of actual location, Naive Bayes classifier is used to calculate the matching probability of current data and the data in the fingerprint database. Finally, according to the principle of path matching positioning method, the current position and motion path were dynamically determined. In this paper, the positioning errors of different positioning models were also verified through experiments, and the positioning accuracy after using Bayes classifier was significantly improved compared with static positioning models of k-weighted neighbor method.

RAS activation is common in human cancers with even few mutations of Ras, indicating alternative regulation leading to aberrant activation of RAS signaling. We identified a Ras GTPase-activating protein gene RASA5/SYNGAP, located at a common 6p21.3 deletion, being methylated and downregulated in several carcinomas, while other RASA family members (RASA1-4) were broadly expressed and only occasionally downregulated in tumors, indicating that RASA5 has a unique and important role in tumorigenesis among RASA family. Unlike other RASA members like RASA1, mutations are rare for RASA5, while its promoter CpG methylation is frequent in cell lines and primary carcinomas of esophageal, nasopharyngeal, breast, lung, gastric and colon, and associated with patient poor survival. We further found that RASA5 inhibited tumor cell migration/invasion, and growth in mouse model, thus as a tumor suppressor. RASA5 suppressed RAS signaling, depending on its RasGAP catalytic activity, which could be counteracted by oncogenic HRas Q61L mutant. Meanwhile, RASA5 knockdown enhanced Ras signaling to promote tumor cell growth. RASA5 also inhibited epithelial-mesenchymal transition through regulating actin remodeling. Thus, our study characterized a novel tumor suppressive role of RASA5, in addition to its known functions in neuro cell regulation. Epigenetic inactivation of RASA5 contributing to hyperactive RAS signaling might have an important role in Ras-driven oncogenesis, particularly in carcinomas with few oncogenic Ras mutations.

Developments towards higher resolution and the ability to detect small defects are bringing a step-change in non-destructive testing. This paper presents a new method for increasing resolution, using a focused electromagnetic acoustic transducer (EMAT) optimised to generate Rayleigh waves at 2 MHz. This high frequency allows detection of mm-depth defects, and the focusing allows sizing of much shorter defects than is\ud possible when using standard EMATs. The focusing behaviour and the aperture angle effect are analysed using laser vibrometry and finite element modelling, showing that a reduced aperture shifts the focal point from the designed value and increases the focal depth. The dual-EMAT has excellent signal to noise ratio (up to 30 dB) and has been used in single shot mode to image a variety of surface-breaking defects, including detecting and positioning a pair of real defects in an aluminium billet sample, and a machined defect of\ud 2 mm length, 0.2 mm width, and 1.5 mm depth, giving an upper limit on the defect length of 2.1 plus or minus 0.5 mm. The results can be used to design an EMAT with optimised focal behaviour for defect detection.

Tumor suppressor p53 prevents cell transformation by inducing apoptosis and other responses. Homozygous TP53 deletion occurs in various types of human cancers for which no therapeutic strategies have yet been reported. TCGA database analysis shows that the TP53 homozygous deletion locus mostly exhibits co-deletion of the neighboring gene FXR2, which belongs to the Fragile X gene family. Here, we demonstrate that inhibition of the remaining family member FXR1 selectively blocks cell proliferation in human cancer cells containing homozygous deletion of both TP53 and FXR2 in a collateral lethality manner. Mechanistically, in addition to its RNA-binding function, FXR1 recruits transcription factor STAT1 or STAT3 to gene promoters at the chromatin interface and regulates transcription thus, at least partially, mediating cell proliferation. Our study anticipates that inhibition of FXR1 is a potential therapeutic approach to targeting human cancers harboring TP53 homozygous deletion., eLife digest Healthy human cells employ many tricks to avoid becoming cancerous. For example, they produce proteins known as tumor suppressors, which sense if a cell shows early signs of cancer and instruct the cell to die. A gene known as TP53 produces one of the most important tumor suppressor proteins, and this gene is inactive or missing in many types of human cancer. Treating cancers that have completely lost the TP53 gene is particularly difficult. One way to develop new treatments for these conditions would be to target other proteins that these cancers need to survive; but these proteins first need to be identified. Fan et al. have now identified one such protein in human cancer cells lacking TP53. Searching databases of DNA sequences from human cancer cells revealed that those without the TP53 gene often also lose a neighboring gene called FXR2. Cancer cells survive without FXR2 because a similar gene, called FXR1, can compensate. Fan et al. therefore decided to experimentally lower the activity of the FXR1 gene and, as expected, cancer cells without TP53 and FXR2 stopped growing. Normal cells, on the other hand, were unaffected by the deletion of the FXR1 gene since FXR2 is still there. This phenomenon, in which cancer cells become vulnerable after the loss of certain genes but only because they have already lost important tumor suppressors, is called “collateral lethality”. Further experiments showed that the protein encoded by FXR1 coordinates with other proteins to activate genes that contribute to cell growth. These findings suggest new ways to treat human cancers that have lost TP53. For example, if scientists can find small molecules that inhibit the protein encoded by FXR1 and show that these molecules can block the growth of tumors lacking TP53 and FXR2, this could eventually lead to a new anticancer drug. However, like any new drug, these small molecule inhibitors would also need to be extensively tested before they could be taken into human clinical trials.

Electromagnetic Acoustic Transducers (EMATs) are a useful ultrasonic tool for non-destructive evaluation in harsh environments due to their non-contact capabilities, and their ability to operate through certain coatings. This work presents a new Rayleigh wave EMAT transducer design, employing geometric focusing to improve the signal strength and detection precision of surface breaking defects. The design is robust and versatile, and can be used at frequencies centered around 1 MHz. Two coils are used in transmission mode, which allows the usage of frequency-based measurement of the defect depth. Using a 2 MHz driving signal, a focused beam spot with a width of 1.3±0.25 mm and a focal depth of 3.7±0.25 mm is measured, allowing for defect length measurements with an accuracy of±0.4 mm and detection of defects as small as 0.5 mm depth and 1 mm length. A set of four coils held under one magnet is used to find defects at orientations offset from normal to the ultrasound beam propagation direction. This EMAT has a range which allows detection of defects which propagate at angles from 16° to 170° relative to the propagation direction over the range of 0–180°, and the setup has the potential to be able to detect defects propagating at all angles relative to the wave propagation direction if two coils are alternately employed as generation coils.

Ultrasonic Rayleigh waves can be employed for the detection of surface breaking defects such as rolling contact fatigue and stress corrosion cracking. Electromagnetic Acoustic Transducers (EMATs) are well suited to this technique as they can directly generate Rayleigh waves within the sample without the requirement for wedges, and they are robust and inexpensive compared to laser ultrasonics. Three different EMAT coil types have been developed, and these are compared to assess their ability to detect and characterize small (down to 0.5 mm depth, 1 mm diameter) surface breaking defects in aluminium. These designs are: a pair of linear meander coils used in a pseudo-pulse-echo mode, a pair of focused meander coils also used in pseudo-pulse-echo mode, and a pair of focused racetrack coils used in pitch-catch mode. The linear meander coils are able to detect most of the defects tested, but have a much lower signal to noise ratio and give limited\ud sizing information. The focused meander coils and the focused racetrack coils can detect all defects tested, but have the advantage that they can also characterize the defect sizes on the sample surface, and have a stronger sensitivity at their focal point. Measurements using all three EMAT designs are presented and compared for high resolution imaging of\ud surface-breaking defects.

An investigation was performed into the effects of operating an absolute eddy-current testing (ECT) probe at frequencies close to its electrical resonance. A previously undocumented defect signal enhancement phenomenon, resulting from associated shifts in electrical resonant frequency, was observed and characterized. Experimental validation was performed on three notch defects on a typical aerospace superalloy, Titanium 6Al–4V. A conventional absolute ECT probe was operated by sweeping through a frequency range about the electrical resonance of the system (1 � 5 MHz). The phenomenon results in signal-to-noise ratio (SNR) peak enhancements by a factor of up to 3.7, at frequencies approaching resonance, compared to those measured at 1 MHz. The defect signal enhancement peaks are shown to be a result of resonant frequency shifts of the system due to the presence of defects within the material. A simple, operational approach for raising the sensitivity of conventional industrial eddycurrent testing is proposed, based on the principles of the observed near electrical resonance signal enhancement (NERSE) phenomenon. The simple procedural change of operating within the NERSE frequency band does not require complex probe design, data analysis or, necessarily, identical coils. Therefore, it is a valuable technique for improving sensitivity, which complements other ECT methods. & 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license

Taurine is the most abundant amino acid in the retina. In the 1970s, it was thought to be involved in retinal diseases with photoreceptor degeneration, because cats on a taurine-free diet presented photoreceptor loss. However, with the exception of its introduction into baby milk and parenteral nutrition, taurine has not yet been incorporated into any commercial treatment with the aim of slowing photoreceptor degeneration. Our recent discovery that taurine depletion is involved in the retinal toxicity of the antiepileptic drug vigabatrin has returned taurine to the limelight in the field of neuroprotection. However, although the retinal toxicity of vigabatrin principally involves a deleterious effect on photoreceptors, retinal ganglion cells (RGCs) are also affected. These findings led us to investigate the possible role of taurine depletion in retinal diseases with RGC degeneration, such as glaucoma and diabetic retinopathy. The major antioxidant properties of taurine may influence disease processes. In addition, the efficacy of taurine is dependent on its uptake into retinal cells, microvascular endothelial cells and the retinal pigment epithelium. Disturbances of retinal vascular perfusion in these retinal diseases may therefore affect the retinal uptake of taurine, resulting in local depletion. The low plasma taurine concentrations observed in diabetic patients may further enhance such local decreases in taurine concentration. We here review the evidence for a role of taurine in retinal ganglion cell survival and studies suggesting that this compound may be involved in the pathophysiology of glaucoma or diabetic retinopathy. Along with other antioxidant molecules, taurine should therefore be seriously reconsidered as a potential treatment for such retinal diseases.

Deletion of 3p12-22 is frequent in multiple cancer types, indicating the presence of critical tumor-suppressor genes (TSGs) at this region. We studied a novel candidate TSG, CMTM7, located at the 3p22.3 CMTM-gene cluster, for its tumor-suppressive functions and related mechanisms. The three CMTM genes, CMTM6, 7 and 8, are broadly expressed in human normal adult tissues and normal epithelial cell lines. Only CMTM7 is frequently silenced or downregulated in esophageal and nasopharyngeal cell lines, but uncommon in other carcinoma cell lines. Immunostaining of tissue microarrays for CMTM7 protein showed its downregulation or absence in esophageal, gastric, pancreatic, liver, lung and cervix tumor tissues. Promoter CpG methylation and loss of heterozygosity were both found contributing to CMTM7 downregulation. Ectopic expression of CMTM7 in carcinoma cells inhibits cell proliferation, motility and tumor formation in nude mice, but not in immortalized normal cells, suggesting a tumor inhibitory role of CMTM7. The tumor-suppressive function of CMTM7 is associated with its role in G1/S cell cycle arrest, through upregulating p27 and downregulating cyclin-dependent kinase 2 (CDK2) and 6 (CDK6). Moreover, CMTM7 could promote epidermal growth factor receptor (EGFR) internalization, and further suppress AKT signaling pathway. Thus, our findings suggest that CMTM7 is a novel 3p22 tumor suppressor regulating G1/S transition and EGFR/AKT signaling during tumor pathogenesis.

Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus-associated tumor prevalent in southern China and southeast Asia, with the 3p14-p12 locus reported as a critical tumor suppressor gene (TSG) region during its pathogenesis. We identified a novel 3p14.2 TSG, FEZF2 (FEZ family zinc finger 2), for NPC. FEZF2 is readily expressed in normal tissues including upper respiratory epithelium, testis, brain and ovary tissues, as well as in immortalized nasopharyngeal epithelial cell line NP69, but it is completely silenced in NPC cell lines due to CpG methylation of its promoter, although no homozygous deletion of FEZF2 was detected. 5-Aza-2'-deoxycytidine treatment restored FEZF2 expression in NPC cell lines along with its promoter demethylation. FEZF2 was frequently downregulated in NPC tumors, with promoter methylation detected in 75.5% of tumors, but only in 7.1% of normal nasopharyngeal tissues. Restored FEZF2 expression suppressed NPC cell clonogenicity through inducing G2/M cell cycle arrest and apoptosis and also inhibited NPC cell migration and stemness. FEZF2 acted as a histone deacetylase-associated repressor downregulating multiple oncogenes including EZH2 and MDM2, through direct binding to their promoters. Concomitantly, overexpression of EZH2 was frequently detected in NPC tumors. Thus, we have identified FEZF2 as a novel 3p14.2 TSG frequently inactivated by promoter methylation in NPC, which functions as a repressor downregulating multiple oncogene expression.

This letter presents a compact dual-mode tri-band bandpass filter by using a short-circuited stub-loaded stepped-impedance resonator (SIR) and a short-circuited stub-loaded uniform impedance resonator. Also, a hairpin SIR geometry is introduced to miniature the size of this filter while maintaining excellent performance. The use of a short-circuited stub at the central point of the hairpin SIR can generate two resonant modes in two passbands. Its equivalent circuit structure is analyzed by using the even-odd mode theory. For demonstration purposes, a tri-band filter for the applications of the Global Positioning System at 1.57 GHz, Worldwide Interoperability for Microwave Access at 3.5 GHz, and wireless local area networks at 5.2 GHz is designed, fabricated, and measured.

// Tingxiu Xiang 1 , Yichao Fan 2 , Chunhong Li 3 , Lili Li 2 , Ying Ying 2 , Junhao Mu 1 , Weiyan Peng 1 , Yixiao Feng 1 , Michael Oberst 4 , Kathleen Kelly 4 , Guosheng Ren 1 , Qian Tao 1, 2 1 Chongqing Key Laboratory of Molecular Oncology and Epigenetics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China 2 Cancer Epigenetics Laboratory, Department of Clinical Oncology, Sir YK Pao Center for Cancer and Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong and CUHK Shenzhen Research Institute, Hong Kong 3 Oncology Department, Suining Sichuan Center Hospital, Sichuan, China 4 Signal Transduction Section, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA Correspondence to: Guosheng Ren, email: rgs726@163.com Qian Tao, email: qtao@cuhk.edu.hk Keywords: DACT2, tumor suppressor, methylation, EMT, breast cancer Received: April 22, 2016 Accepted: September 14, 2016 Published: September 29, 2016 ABSTRACT Wnt signaling plays an important role in breast carcinogenesis. DAPPER2 (DACT2) functions as an inhibitor of canonical Wnt signaling and plays distinct roles in different cell contexts, with its role in breast tumorigenesis unclear. We investigated DACT2 expression in breast cancer cell lines and primary tumors, as well as its functions and molecular mechanisms. Results showed that DACT2 expression was silenced in 9/9 of cell lines. Promoter CpG methylation of DACT2 was detected in 89% (8/9) of cell lines, as well as in 73% (107/147) of primary tumors, but only in 20% (1/5) of surgical margin tissues and in none of normal breast tissues. Demethylation of BT549 and T47D cell lines with 5-aza-2’-deoxycytidine restored DACT2 expression along with promoter demethylation, suggesting that its downregulation in breast cancer is dependent on promoter methylation. Furthermore, ectopic expression of DACT2 induced breast cell apoptosis in vitro , and further inhibited breast tumor cell proliferation, migration and EMT, through antagonizing Wnt/β-catenin and Akt/GSK-3 signaling. Thus, these results demonstrate that DACT2 functions as a tumor suppressor for breast cancer but was frequently disrupted epigenetically in this cancer.

The sensitivity enhancing effects of eddy-current testing at frequencies close to electrical resonance are explored. Var-ied techniques exploiting the phenomenon, dubbed near electrical resonance signal enhancement (NERSE), were experimentally investigated to evaluate its potential exploitation for other interesting applications in aerospace materials, in particular its potential for boosting the sensitivity of standard ECT measurements. Methods for setting and controlling the typically unstable resonant frequencies of such systems are discussed. This research is funded by the EPSRC, via the Research Centre for Non-Destructive Evaluation RCNDE, and Rolls-Royce plc.

B-lymphocyte-induced maturation protein 1 (BLIMP1) exists as two major isoforms, α and β, which arise from alternate promoters. Inactivation of the full length BLIMP1α isoform is thought to contribute to B cell lymphomagenesis by blocking post-germinal centre (GC) B cell differentiation. In contrast, the shorter β isoform is functionally impaired and over-expressed in several haematological malignancies, including diffuse large B cell lymphomas (DLBCL). We have studied the influence on BLIMP1β expression of the Epstein-Barr virus (EBV), a human herpesvirus that is implicated in the pathogenesis of several GC-derived lymphomas, including a subset of DLBCL and Hodgkin’s lymphoma (HL). We show that BLIMP1β expression is increased following the EBV infection of normal human tonsillar GC B cells. We also show that this change in expression is accompanied by hypomethylation of the BLIMP1β-specific promoter. Furthermore, we confirmed previous reports that the BLIMP1β promoter is hypomethylated in DLBCL cell lines and show for the first time that BLIMP1β is hypomethylated in the Hodgkin/Reed-Sternberg (HRS) cells of HL. Our results provide evidence in support of a role for BLIMP1β in the pathogenesis of EBV-associated B cell lymphomas.

Beta1, 4-Galactosyltransferase-I (β1, 4-GalT-I), which transfers galactose from UDP-Gal to N-acetylglucosamine and N-acetylglucosamine-terminated oligosaccharides of N- and O-linked glycans in a β(1-4) linkage, plays a critical role in cell adhesion, sperm-egg recognition, neurite growth, and tumor cell migration and invasion. Our previously experiments also show that β1, 4-GalT-I was up-regulated by estrogens and some important cytokines of embryo implantation especially Interleukin-1 (IL-1), TGF-α and Leukemia Inhibitory Factor (LIF) in endometrial cells. In the receptive phase human uterus, osteopontin (OPN) is the most highly up-regulated extracellular matrix/adhesion molecule/cytokine. In this study, we demonstrated the correlated expression of OPN and β1, 4-GalT-I in endometrium during early pregnancy, and recombinant human OPN (rhOPN) protein induced the β1, 4-GalT-I up-regulation in RL95-2 cells. Inhibition of MEK/ERK, PI3K/AKT and NF-κB suppressed rhOPN-induced β1, 4-GalT-I expression. In addition, rhOPN promoted the adhesion of blastocysts cells in vitro in β1, 4-GalT-I-dependent manner. Moreover, the adhesion is greatly inhibited when β1, 4-GalT-I was blocked with the specific antibody. Taken together, our data suggest that β1, 4-GalT-I provides a mechanism to bridge embryo to endometrium during implantation.

The malignant Hodgkin/Reed-Sternberg (HRS) cells of Hodgkin lymphoma (HL) are believed to derive from germinal center (GC) B cells, but lack expression of a functional B cell receptor. As apoptosis is the normal fate of B-cell receptor–negative GC B cells, mechanisms that abrogate apoptosis are thus critical in HL development, such as epigenetic disruption of certain pro-apoptotic cancer genes including tumor suppressor genes. Identifying methylated genes elucidates oncogenic mechanisms and provides valuable biomarkers; therefore, we performed a chemical epigenetic screening for methylated genes in HL through pharmacological demethylation and expression profiling. IGSF4/CADM1/TSLC1, a pro-apoptotic cell adhesion molecule of the immunoglobulin superfamily, was identified together with other methylated targets. In contrast to its expression in normal GC B cells, IGSF4 was down-regulated and methylated in HL cell lines, most primary HL, and microdissected HRS cells of 3/5 cases, but not in normal peripheral blood mononuclear cells and seldom in normal lymph nodes. We also detected IGSF4 methylation in sera of 14/18 (78%) HL patients but seldom in normal sera. Ectopic IGSF4 expression decreased HL cells survival and increased their sensitivity to apoptosis. IGSF4 induction that normally follows heat shock stress treatment was also abrogated in methylated lymphoma cells. Thus, our data demonstrate that IGSF4 silencing by CpG methylation provides an anti-apoptotic signal to HRS cells important in HL pathogenesis.

On the UK rail network, changing usage patterns have led to rolling contact fatigue (RCF) on the railhead being an increasing problem and frequent testing for such defects is essential. Existing systems for detecting defects are limited in speed ( 5 mm). We are currently working on novel ultrasonic methods of detecting RCF, designed to lift many of the speed limitations and/ or significantly improve accuracy and resolution. A noncontact pitch-catch pulsed ultrasonic system comprising two electromagnetic acoustic transducers (EMATs) generating and detecting Rayleigh-like waves has the potential to operate at high speed, thus improving network inspection coverage. Phased array ultrasonic techniques are being designed to improve imaging and characterisation of the defects. A set of simulated defects on sections of rail have been machined, including half-face slots machined normal to the railhead surface, clusters of angled slots, and pocket defects more typical of real defects. The techniques have been tested on these rails and their potential for use on the rail network is discussed.

Electromagnetic acoustic transducers (EMATs) are non-contact ultrasonic transducers capable of generating wide band ultrasonic surface waves on metallic samples. A lab-based laser-EMAT system has been developed to observe the ultrasonic surface wave propagation and interaction with surface breaking defects on the sample rail head surface. A wide band EMAT generating surface waves with a frequency content between approximately 50 and 500 kHz is used to propagate ultrasonic waves on the surface of a rail head down the length of the sample. A stabilised Michelson interferometer is used to measure the out-of-plane displacement of the surface wave. A complete picture of the ultrasonic surface wave on the sample surface over time is reconstructed using this technique, with exceptionally high spatial and temporal resolution. Despite the curvature of the rail head, the ultrasonic surface wave propagating down the rail is found to have similar properties to Rayleigh waves by direct comparison to those observed on flat samples using the same technique.

Aims: Nasopharyngeal carcinoma (NPC) is a common tumor consistently associated with Epstein–Barr virus infection and prevalent in South China, including Hong Kong, and southeast Asia. Current genomic sequencing studies found only rare mutations in NPC, indicating its critical epigenetic etiology, while no epigenome exists for NPC as yet. Materials & methods: We profiled the methylomes of NPC cell lines and primary tumors, together with normal nasopharyngeal epithelial cells, using methylated DNA immunoprecipitation (MeDIP). Results: We observed extensive, genome-wide methylation of cellular genes. Epigenetic disruption of Wnt, MAPK, TGF-β and Hedgehog signaling pathways was detected. Methylation of Wnt signaling regulators (SFRP1, 2, 4 and 5, DACT2, DKK2 and DKK3) was frequently detected in tumor and nasal swab samples from NPC patients. Functional studies showed that these genes are bona fide tumor-suppressor genes for NPC. Conclusion: The NPC methylome shows a special high-degree CpG methylation epigenotype, similar to the Epstein–Barr virus-infected gastric cancer, indicating a critical epigenetic etiology for NPC pathogenesis.

Purpose Exendin-4 (E4), a long-acting agonist of the hormone glucagon-like peptide 1 receptor (GLP-1R), is administered to treat type II diabetes in the clinical setting and also shows a neuroprotective effect. Our previous studies demonstrated its protective effect in early experimental diabetic retinopathy (DR), but the molecular and cellular mechanisms are largely unknown. This study aimed to investigate the protective mechanism of a GLP-1R agonist E4 against early DR in Goto-Kakizaki (GK) rats. Methods Diabetic GK rats and control animals were randomly assigned to receive E4 or vehicle by intravitreal injection. The retinal function and retinal cell counts were evaluated using an electroretinogram and light microscopy. The expressions of retinal GLP-1R, mitochondria-dependent apoptosis-associated genes, reactive gliosis markers, and endoplasmic reticulum stress-related pathway genes were studied by western blotting and immunohistochemistry in vivo and in vitro. Results E4 significantly prevented the reduction of the b-wave and oscillatory potential amplitudes and retinal cell loss and maintained the Bcl-2/Bax and Bcl-xL/Bax ratio balances in GK rats. It also downregulated the expression of glial fibrillary acidic protein and reduced retinal reactive gliosis. Similar results were found in primary rat Müller cells under high glucose culture in vitro. Conclusions E4 may protect retinal cells from diabetic attacks by activating GLP-1R, decreasing retinal cell apoptosis, and reducing retinal reactive gliosis. Thus, E4 treatment may be a novel approach for early DR.

We present simultaneous measurements of acoustic emission and magnetic Barkhausen noise during the thermally induced martensitic transition in a Ni-Mn-Ga single crystal. The range where structural acoustic emission avalanches are detected extends for more than 50 K for both cooling and heating ramps, with a hysteresis of ∼10 K. The magnetic activity occurs during the structural transition, exhibiting similar hysteresis, but concentrated in the lower half of the temperature range. Statistical analysis of individual signals allows characterization of the broad distributions of acoustic emission and Barkhausen amplitudes. By studying the times of arrival of the avalanche events we detect the existence of correlations between the two kinds of signals, with a number of acoustic emission signals occurring shortly after a Barkhausen signal. The order of magnitude of the observed delays is compatible with the time needed for the propagation of ultrasound through the sample, showing correlation of some of the signals.

The use of phased array methods are commonplace in ultrasonic applications, where controlling the variation of the phase between the narrowband emitters in an array facilitates beam steering and focusing of ultrasonic waves. An approach is presented here whereby emitters of alternating polarity arranged in a one-dimensional array are pulsed simultaneously, and have sufficiently wide, controlled bandwidth to emit a two-dimensional wave. This pulsed approach provides a rapid means of simultaneously covering a region of space with a wave-front, whereby any wave that scatters or reflects off a body to a detector will have a distinct arrival time and frequency. This is a general wave phenomenon with a potential application in radar, sonar, and ultrasound. The key result is that one can obtain a smooth, continuous wave-front emitted from the array, over a large solid angle, whose frequency varies as a function of angle to the array. Analytic and finite element models created to describe this phenomenon have been validated with experimental results using ultrasonic waves in metal samples.

Conventional ultrasonic mensuration of sample thickness from one side only requires the bulk wave reverberation time and a calibration speed. This speed changes with temperature, stress, and microstructure, limiting thickness measurement accuracy. Often, only one side of a sample is accessible, making in situ calibration impossible. Non-contact ultrasound can generate multiple shear horizontal guided wave modes on one side of a metal plate. Measuring propagation times of each mode at different transducer separations, allows sheet thickness to be calculated to better than 1% accuracy for sheets of at least 1.5mm thickness, without any calibration. (Some figures may appear in colour only in the online journal)

Three designs for electrodynamic flexural transducers (EDFT) for air-coupled ultrasonics are presented and compared. An all-metal housing was used for robustness, which makes the designs more suitable for industrial applications. The housing is designed such that there is a thin metal plate at the front, with a fundamental flexural vibration mode at ∼50 kHz. By using a flexural resonance mode, good coupling to the load medium was achieved without the use of matching layers. The front radiating plate is actuated electrodynamically by a spiral coil inside the transducer, which produces an induced magnetic field when an AC current is applied to it. The transducers operate without the use of piezoelectric materials, which can simplify manufacturing and prolong the lifetime of the transducers, as well as open up possibilities for high-temperature applications. The results show that different designs perform best for the generation and reception of ultrasound. All three designs produced large acoustic pressure outputs, with a recorded sound pressure level (SPL) above 120 dB at a 40 cm distance from the highest output transducer. The sensitivity of the transducers was low, however, with single shot signal-to-noise ratio ( SNR ) ≃ 15 dB in transmit–receive mode, with transmitter and receiver 40 cm apart.

Electromagnetic acoustic transducers (EMATs) have been used in pitch-catch mode for a better characterization of surface cracks in metals. The system, which combines the measurement of Rayleigh wave velocity in the in-plane and out-of-plane directions, has been used to understand the interaction of Rayleigh waves with inclined surface cracks. This shows a stronger and more prominent enhancement pattern when compared to the enhancement caused by cracks which are normal to the surface. In addition, measurements in the far-field are combined with the near-field enhancement measurement, with a view to characterize the cracks. An algorithm for characterizing surface cracks is presented. A finite element method model has been computed to simulate the experiment, and the cause of the prominent enhancement in the inclined cracks is explained.

A pulsed Nd: YAG laser with an approximately Gaussian beam shape is directed onto the surface of an aluminium sheet at an energy density below which damage by laser ablation occurs, generating Lamb waves in the sheet. The laser beam is raster scanned across the surface of the sample. The Lamb waves travel radially outwards from the generation point and are detected some distance away by an electromagnetic acoustic transducer with sensitivity to in-plane displacements of the sheet. A number of static EMATs are located around the edges of the sheet, some distance from the generation point. The presence of a crack-like defect on the sheet can be detected by either a sudden change in the ultrasonic waveform or by an enhancement in the frequency content of the waveform when the laser beam illuminates directly onto the crack.

Chromosome 3p harbors multiple tumor-suppressor genes. PLCD1, located at 3p22, encodes an enzyme that mediates regulatory signaling of energy metabolism, calcium homeostasis and intracellular movement. We investigated the epigenetic alterations of PLCD1 and its tumor suppressor function in breast cancer. Frequent downregulation/silencing of PLCD1 was shown in most breast cancer cell lines by using semi-quantitative PCR. Promoter methylation of PLCD1 was detected in 78% (7/9) of cell lines and 52% (13/25) of primary tumors by Methylation-specific PCR (MSP), but not in any tumor adjacent breast tissues and normal breast tissues, which was further confirmed by bisulfite genomic sequencing (BGS). The silencing of PLCD1 could be reversed by pharmacological demethylation, indicating a methylation-mediated mechanism. Ectopic expression of PLCD1 in silenced breast cancer cells significantly inhibited their colony formation. In addition, PLCD1 inhibited tumor cell migration and induced cell cycle G(2)/M arrest. Thus, this study for the first time demonstrates the frequent inactivation of PLCD1 by promoter methylation and its tumor inhibitory function in breast cancer. Tumor-specific methylation of PLCD1 might serve as a biomarker for possible early detection and prognosis prediction of breast cancer.

WNT5A is classified as a non-transforming WNT family member whose role in carcinogenesis is still ambiguous. It exhibits tumor suppressor activities in some cancers (thyroid, brain, breast and colorectum), but is aberrantly upregulated in cancers of lung, stomach and prostate. We investigated its epigenetic alterations and functions in esophageal squamous cell carcinoma (ESCC). With semi-quantitative reverse transcription PCR, we found that WNT5A was silenced or downregulated in 5 of 18 ESCC cell lines, but expressed in normal esophagus tissue and immortalized normal esophageal epithelial cell lines. Promoter CpG methylation of WNT5A was detected in 4 of the 5 downregulated ESCC cell lines, while 5-aza-2'-deoxycytidine treatment induced WNT5A expression and demethylated its promoter in silenced cell lines. WNT5A promoter methylation was frequently detected in primary ESCC (24/36, 67%), but less frequently and weakly in paired surgical marginal esophageal tissues (8/36, 22%; p0.01), while no methylation was detected in seven normal esophageal epithelial tissues from healthy donors. Ectopic expression of WNT5A resulted in significant inhibition of clonogenicity and motility of ESCC cells, accompanied by a dramatic decrease of intracellular β-catenin protein level and β-catenin transcriptional activity. In summary, we show that WNT5A is frequently silenced in ESCC by promoter methylation and exhibits tumor suppressor properties through antagonizing the WNT/β-catenin pathway. The epigenetic disruption of WNT5A would thus contribute directly to the aberrant activation of WNT/β-catenin signaling during ESCC pathogenesis.

Identifying tumor suppressor genes silenced by promoter CpG methylation uncovers mechanisms of tumorigenesis and identifies new epigenetic biomarkers for early cancer detection. DLEC1 is located at 3p22.3, a critical tumor suppressor gene locus for renal cell carcinoma. We explored its epigenetic alteration in renal cell carcinoma and possible clinicopathological association.We examined DLEC1 expression and methylation by semiquantitative reverse transcriptase and methylation specific polymerase chain reaction in 9 renal cell carcinoma cell lines and 81 primary tumors. We also analyzed the relationship between DLEC1 methylation and clinicopathological features in patients with renal cell carcinoma. We assessed DLEC1 inhibition of renal cell carcinoma cell growth by colony formation assay.DLEC1 methylation and down-regulation were detected in all renal cell carcinoma cell lines. Treatment with 5-aza-2'-deoxycytidine (Sigma) and/or trichostatin A (Cayman Chemical, Ann Arbor, Michigan) reversed methylation and restored DLEC1 expression, indicating that methylation directly mediates its silencing. Aberrant methylation was further detected in 25 of 81 primary tumors (31%) but only 1 of 53 nonmalignant renal tissues (2%) showed methylation. DLEC1 methylation status was significantly associated with TNM classification and grade in patients with renal cell carcinoma (chi-square test p = 0.01 and 0.04, respectively). DLEC1 ectopic expression in silenced renal cell carcinoma cells resulted in substantial tumor cell clonogenicity inhibition.To our knowledge we report for the first time that DLEC1 is often down-regulated by CpG methylation and shows tumor inhibitory function in renal cell carcinoma cells, indicating its role as a tumor suppressor. DLEC1 tumor specific methylation may serve as a biomarker for early detection and prognosis prediction of this tumor.

Laser generated ultrasound has been widely used for detecting cracks, surface and sub-surface defects in\ud many different materials. It provides a non-contact wideband excitation source which can be focused into\ud different geometries. Previous workers have reported enhancement of the laser generated Rayleigh wave\ud when a crack is illuminated by pulsed laser beam irradiation. We demonstrate that the enhancement\ud observed is due to a combination of source truncation, the free boundary condition at the edge of the crack\ud and interference effects. Generating a Rayleigh wave over a crack can lead to enhancement of the\ud amplitude of the Rayleigh wave signal, a shift in the dominant frequency of the wideband Rayleigh wave\ud and strong enhancement of the high frequency components of the Rayleigh wave.